A kind of tissue clearing liquid sut and its preparation and application

Abstract

An embodiment of the invention relates to a tissue vitrification liquid SUT, which mainly comprises SDS, urea and TritonX-100. The invention further relates to a preparation and application of the tissue vitrification liquid SUT. The tissue vitrification liquid SUT provided in the embodiment is reduced in preparation cost and difficulty, is improved in vitrification effect, and has a cardiac muscle vitrification degree superior than that of some vitrification liquids in the prior art.

Description

technical field [0001] The invention relates to a tissue clearing fluid SUT and its preparation and application. Background technique [0002] It has always been a dream of scientists to obtain visualized and complete tissue structure information at the single-cell resolution level. Through single-cell resolution imaging of intact tissues, three-dimensional stereoscopic images of intact tissues can be obtained, so as to clarify the connection mode and dynamic changes between cells at the organ and system level, which will greatly improve our understanding of the occurrence and development of diseases Recognize that single-cell events in disease can ultimately lead to changes in the health of the entire organism. But the opacity of tissues and organs has become the biggest obstacle to realize this dream. [0003] Classical protein qualitative and quantitative techniques (that is, to determine which protein is the content of a certain protein) include western blot, immunohis...

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Problems solved by technology

However, the birth of various methods is mainly used for the study of the nervous system (brain tissue), and 3DISCO has been used for other organs to be transparent, but it is easy to cause fluorescence quenching

The clearing methods published in the journal CELL in 2014 include CUBIC and PACT-PARS, in which the clearing solution of CUBIC contains two reagents, which need two reagents to treat the tissue in sequence; the clearing solution of PACT-PARS is 8% SDS, two The whole body of the mouse was cleared by perfusing the two clearing fluids through the circulatory system. PACT-PARS also performed confocal scanning imaging of the brain, which vividly displayed the structure of brain neurons. However, research on other organs still remains to be done. superficial

[0008] (1) Although the transparency effect of CUBIC is good, the composition of the reagents is complex and involves uncommon reagents. Among them, N,N,N',N'-tetrakis(2-hydroxypropyl)ethylenediamine (ie EDTP) is very viscous. It is difficult to prepare the solution, and the clearing reagent contains two kinds, which need to be used sequentially. The processing time of each reagent is different, and the operation steps are relatively complicated. Moreover, since EDTP is a copper ion sedimentation agent, the conventional BCA method cannot be used for protein quantification. At the same time, because CUBIC-1 is relatively viscous, protein quantification with Nanodrop is not accurate;

[0009] (2) 8% SDS is simple, but the effect of clearing is poor, and the tissue swelling is serious, and the protein loss is more

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